Abstract: Objective To detect UGT1A1 gene polymorphism for neonatal patients with high blood bilirubin by using multiple real-time fluorescent quantitative PCR technique and to discuss the relationship between UGT1A1 gene polymorphism and neonatal bilirubin level. Methods We chose 165 patients with high neonatal bilirubin（bilirubin≥257μmol/L） as the test group and 150 general subjects(bilirubin<257μmol/L) as control group in authors’hospitals between January 2015 and January 2016. DNA was extracted firstly from the whole blood;four common mutations of UGT1A1 gene were chosen,including A(TA)7TAA box of the promoter,and the Pro229Gln,Try486Asp,Gly71Arg in the coding region;primers and probe were synthesized;and samples were detected by multiple real-time fluorescent quantitative PCR technique and the fluorescence was collected;and the samples with positive results were recovered and purified for gene sequencing. Results The results of multiple real-time fluorescent quantitative PCR were consistent with the results of gene sequencing. The detection rates of A(TA)7TAA box mutation in test group and control group were 7.8% and 23.3% respectively,while the rate of Gly71Arg mutation in the test group and control group were 52.7% and 19.3% respectively,and the difference was statistically significant. However,the detection rates of Try486Asp and Pro229Gln mutation were relatively low with no statistical significance. In the test group,the bilirubin level of patients with homozygous Gly71Arg was higher than that of wild type or heterozygous mutations,and the difference was statistically significant. Conclusion Multiple real-time fluorescent quantitative PCR technique is a convenient, accurate,efficient and economic way to detect gene mutation,and the neonatal hyperbilirubinemia is associated with Gly71Arg mutations.

Key words: Multiple real-time fluorescent quantitative PCR technique, Nenatal hyperbilirubinemia